Regulation of vascular endothelial (VE) growth factor (VEGF)-induced permeability is critical in physiological and pathological processes. We show that tyrosine phosphorylation of VEGF receptor 2 ...(VEGFR2) at Y951 facilitates binding of VEGFR2 to the Rous sarcoma (Src) homology 2-domain of T cell-specific adaptor (TSAd), which in turn regulates VEGF-induced activation of the c-Src tyrosine kinase and vascular permeability. c-Src was activated in vivo and in vitro in a VEGF/TSAd-dependent manner, and was regulated via increased phosphorylation at pY418 and reduced phosphorylation at pY527. Tsad silencing blocked VEGF-induced c-Src activation, but did not affect pathways involving phospholipase Cγ, extracellular regulated kinase, and endothelial nitric oxide. VEGF-induced rearrangement of VE-cadherin-positive junctions in endothelial cells isolated from mouse lungs, or in mouse cremaster vessels, was dependent on TSAd expression, and TSAd formed a complex with VE-cadherin, VEGFR2, and c-Src at endothelial junctions. Vessels in tsad(-/-) mice showed undisturbed flow and pressure, but impaired VEGF-induced permeability, as measured by extravasation of Evans blue, dextran, and microspheres in the skin and the trachea. Histamine-induced extravasation was not affected by TSAd deficiency. We conclude that TSAd is required for VEGF-induced, c-Src-mediated regulation of endothelial cell junctions and for vascular permeability.
Hand, foot, and mouth disease (HFMD) has been one of the most common infectious diseases in Shijiazhuang City, as is the situation in China overall. In the National HFMD surveillance system, the ...pathogen detection was focused on EV-A71 and CVA16, and therefore, information on the other EVs is very limited. In order to identify the circulating EV serotypes in the HFMD outbreaks in Shijiazhuang City during 2010-2012, 4045 patients presented with HFMD were recruited in the study, and clinical samples were investigated. Typing of EV serotypes was performed using the molecular typing methods, and phylogenetic analyses based on entire VP1 sequences of human enterovirus 71 (EV-A71), coxsackievirus A16 (CVA16), CVA10 and CVB3 was performed. The results revealed that EV-A71 and CVA16 were the 2 most important pathogens but the circulating trends of the 2 viruses showed a shift, the spread of EV-A71 became increasingly weak, whereas the spread of CVA16 became increasingly stronger. CVA10 and CVB3 were the third and fourth most prevalent pathogens, respectively. Co-infection of two viruses at the same time was not found in these samples. Based on entire VP1 region sequences, the phylogenetic analysis revealed that C4a subgenotype EV-A71, B1a and B1b subgenotype CVA16 continued to evolve. The CVA10 strains were assigned to 4 genotypes (A-D), whereas the CVB3 strains were assigned to 5 genotypes (A-E), with clear geographical and temporal-specific distributions. The Shijiazhuang CVA10 sequences belonged to 4 epidemic lineages within genotype C, whereas the Shijiazhuang CVB3 sequences belonged to 2 epidemic lineages within genotype E, which may have the same origins as the strains reported in other part of China. CVA10 and CVB3, 2 pathogens that were previously infrequently detected, were identified as pathogens causing the HFMD outbreaks. This study underscores the need for detailed laboratory-based surveillances of HFMD in mainland China.
The remote sensing technology of unmanned aerial vehicle (UAV) is a low altitude remote sensing technology. The technology has been widely used in military, agricultural, medical, geographical ...mapping, and other fields by virtue of the advantages of fast acquisition, high resolution, low cost, and good security. But limited by the flying height of UAV and the focal length of the digital camera, the single image obtained by the UAV is difficult to form the overall cognition of the ground farmland area. In order to further expand the field of view, it is necessary to mosaic multiple single images acquired by UAV into a complete panoramic image of the farmland. In this paper, aiming at the problem of UAV low-altitude remote sensing image splicing, an image mosaic technique based on Speed Up Robust Feature (SURF) is introduced to achieve rapid image splicing. One hundred fifty ground farmland remote sensing images collected by UAV are used as experimental splicing objects, and the image splicing is completed by the global stitching strategy optimized by Levenberg-Marquardt (L-M). Experiments show that the strategy can effectively reduce the influence of cumulative errors and achieve automatic panoramic mosaic of the survey area.
MicroRNAs (miRNAs) have been implicated to play a central role in the development of drug resistance in a variety of malignancies. However, many studies were conducted at the in vitro level and could ...not provide the in vivo information on the functions of miRNAs in the anticancer drug resistance. Here, we introduced a dual reporter gene imaging system for noninvasively monitoring the kinetic expression of miRNA-16 during chemoresistance in gastric cancer both in vitro and in vivo. Human sodium iodide symporter (hNIS) and firefly luciferase (Fluc) genes were linked to form hNIS/Fluc double fusion reporter gene and then generate human gastric cancer cell line NF-3xmir16 and its multidrug resistance cell line NF-3xmir16/VCR. Radioiodide uptake and Fluc luminescence signals in vitro correlated well with viable cell numbers. The luciferase activities and radioiodide uptake in NF-3xmir16 cells were remarkably repressed by exogenous or endogenous miRNA-16. The NF-3xmir16/VCR cells showed a significant increase of (131)I uptake and luminescence intensity compared to NF-3xmir16 cells. The radioactivity from in vivo (99m)Tc-pertechnetate imaging and the intensity from bioluminescence imaging were also increased in NF-3xmir16/VCR compared with that in NF-3xmir16 tumor xenografts. Furthermore, using this reporter gene system, we found that etoposide (VP-16) and 5-fluorouracil (5-FU) activated miRNA-16 expression in vitro and in vivo, and the upregulation of miRNA-16 is p38MAPK dependent but NF-κB independent. This dual imaging reporter gene may be served as a novel tool for in vivo imaging of microRNAs in the chemoresistance of cancers, as well as for early detection and diagnosis in clinic.
Efficacy of immunotherapy in hepatocellular carcinoma (HCC) is blocked by its high degree of inter‐ and intra‐tumor heterogeneity and immunosuppressive tumor microenvironment. However, the ...correlation between tumor heterogeneity and immunosuppressive microenvironment in HCC has not been well addressed. Here, we endeavored to dissect inter‐ and intra‐tumor heterogeneity in HCC and uncover how they contribute to the immunosuppressive microenvironment. We performed consensus molecular subtyping with non‐negative matrix factorization (NMF) clustering to stratify the inter‐heterogeneity profile of HCC tumors. We grouped HCC tumors from the Cancer Genome Atlas (TCGA) patients into three subtypes (S1, S2 and S3), where S1 was characterized as a ‘hot tumor’ profile with high expression level of T cell genes and rate of immune scores. S2 was characterized as a ‘cold tumor’ profile with the highest tumor purity score, and S3 as an ‘immunosuppressed tumor’ profile with the poorest prognosis and a high expression level of immunosuppressive genes such as cytotoxic T‐lymphocyte‐associated protein‐4, TIGIT, and PDCD1. Moreover, we combined weighted gene co‐expression network analysis and single‐cell regulatory network inference and clustering (SCENIC) in the single‐cell dataset of the S3‐like subtype (CS3) and identified a transcription factor, BATF, which could upregulate immunosuppressive genes. Finally, we identified a cell interaction network in which a myeloid‐derived suppressor cell‐like macrophage subtype could promote the formation of immunosuppressive T‐cells.
By integrating bulk and single‐cell RNA sequencing in this study, we defined a 108‐gene classifier and investigated cellular heterogeneity, as well as immune infiltration in hepatocellular carcinoma (HCC). Patients with a high level of cytotoxic T‐lymphocyte‐associated protein‐4, TIGIT and LAG3 showed the poorest prognosis in both the Cancer Genome Atlas and International Cancer Genome Consortium cohorts. Additionally, single‐cell RNA sequencing analysis revealed high expression levels of these immunosuppressive genes and poor patient survival within the CS3 cluster. Our research indicated that BATF could be regulating the expression of immunosuppressive genes in HCC.
The specific role of VEGFA-induced permeability and vascular leakage in physiology and pathology has remained unclear. Here we show that VEGFA-induced vascular leakage depends on signalling initiated ...via the VEGFR2 phosphosite Y949, regulating dynamic c-Src and VE-cadherin phosphorylation. Abolished Y949 signalling in the mouse mutant Vegfr2(Y949F/Y949F) leads to VEGFA-resistant endothelial adherens junctions and a block in molecular extravasation. Vessels in Vegfr2(Y949F/Y949F) mice remain sensitive to inflammatory cytokines, and vascular morphology, blood pressure and flow parameters are normal. Tumour-bearing Vegfr2(Y949F/Y949F) mice display reduced vascular leakage and oedema, improved response to chemotherapy and, importantly, reduced metastatic spread. The inflammatory infiltration in the tumour micro-environment is unaffected. Blocking VEGFA-induced disassembly of endothelial junctions, thereby suppressing tumour oedema and metastatic spread, may be preferable to full vascular suppression in the treatment of certain cancer forms.
Over-exploitation of coal mines leads to surface subsidence, surface cracks, collapses, landslides, and other geological disasters. Taking a mining area in Nalintaohai Town, Ejin Horo Banner, Ordos ...City, Inner Mongolia Autonomous Region, as an example, Sentinel-1A data from January 2018 to October 2019 were used as the data source in this study. Based on the high interference coherence of the permanent scatterer (PS) over a long period of time, the problem of the manual selection of ground control points (GCPs) affecting the monitoring results during refinement and re-flattening is solved. A DInSAR-PS-Stacking method combining the PS three-threshold method (the coherence coefficient threshold, amplitude dispersion index threshold, and deformation velocity interval) is proposed as a means to select ground control points for refinement and re-flattening, as well as a means to obtain time-series deformation by weighted stacking processing. A SBAS-PS-InSAR method combining the PS three-threshold method to select PS points as GCPs for refinement and re-flattening is also proposed. The surface deformation results monitored by the DInSAR-PS-Stacking and SBAS-PS-InSAR methods are analyzed and verified. The results show that the subsidence location, range, distribution, and space–time subsidence law of surface deformation results obtained by DInSAR-PS-Stacking, SBAS-PS-InSAR, and GPS methods are basically the same. The deformation results obtained by these two InSAR methods have a good correlation with the GPS monitoring results, and the MAE and RMSE are within the acceptable range. The error showed that the edge of the subsidence basin was small and that the center was large. Both methods were found to be able to effectively monitor the coal mine, but there were also shortcomings. DInSAR-PS-Stacking has a strong ability to monitor the settlement center. SBAS-PS-InSAR performed well in monitoring slow and small deformations, but its monitoring of the settlement center was insufficient. Considering the advantages of these two InSAR methods, we proposed fusing the time-series deformation results obtained using these two InSAR methods to allow for more reliable deformation results and to carry out settlement analysis. The results showed that the automatic two-threshold (deformation threshold and average coherence threshold) fusion was effective for monitoring and analysis, and the deformation monitoring results are in good agreement with the actual situation. The deformation information obtained by the comparison, and fusion of multiple methods can allow for better monitoring and analysis of the mining area surface deformation, and can also provide a scientific reference for mining subsidence control and early disaster warning.
Peritoneal fibrosis is the prevailing complication induced by prolonged exposure to high glucose in patients undergoing peritoneal dialysis.
To elucidate the molecular mechanisms underlying this ...process, we conducted an integrated analysis of the transcriptome and chromatin accessibility profiles of human peritoneal mesothelial cells (HMrSV5) during high-glucose treatment.
Our study identified 2775 differentially expressed genes (DEGs) related to high glucose-triggered pathological changes, including 1164 upregulated and 1611 downregulated genes. Genome-wide DEGs and network analysis revealed enrichment in the epithelial-mesenchymal transition (EMT), inflammatory response, hypoxia, and TGF-beta pathways. The enriched genes included VEGFA, HIF-1α, TGF-β1, EGF, TWIST2, and SNAI2. Using ATAC-seq, we identified 942 hyper (higher ATAC-seq signal in high glucose-treated HMrSV5 cells than in control cells) and 714 hypo (lower ATAC-seq signal in high glucose-treated HMrSV5 cells versus control cells) peaks with differential accessibility in high glucose-treated HMrSV5 cells versus controls. These differentially accessible regions were positively correlated (R = 0.934) with the nearest DEGs. These genes were associated with 566 up- and 398 downregulated genes, including SNAI2, TGF-β1, HIF-1α, FGF2, VEGFA, and VEGFC, which are involved in critical pathways identified by transcriptome analysis. Integrated ATAC-seq and RNA-seq analysis also revealed key transcription factors (TFs), such as HIF-1α, ARNTL, ELF1, SMAD3 and XBP1. Importantly, we demonstrated that HIF-1α is involved in the regulation of several key genes associated with EMT and the TGF-beta pathway. Notably, we predicted and experimentally validated that HIF-1α can exacerbate the expression of TGF-β1 in a high glucose-dependent manner, revealing a novel role of HIF-1α in high glucose-induced pathological changes in human peritoneal mesothelial cells (HPMCs).
In summary, our study provides a comprehensive view of the role of transcriptome deregulation and chromosome accessibility alterations in high glucose-induced pathological fibrotic changes in HPMCs. This analysis identified hub genes, signaling pathways, and key transcription factors involved in peritoneal fibrosis and highlighted the novel glucose-dependent regulation of TGF-β1 by HIF-1α. This integrated approach has offered a deeper understanding of the pathogenesis of peritoneal fibrosis and has indicated potential therapeutic targets for intervention.
Among the bioactive compounds, lipid-soluble tanshinone is present in
, a medicinal plant species. While it is known that ethephon has the ability to inhibit the tanshinones biosynthesis in the
hairy ...root, however the underlying regulatory mechanism remains obscure. In this study, using the transcriptome dataset of the
hairy root induced by ethephon, an ethylene-responsive transcriptional factor EIN3-like 1 (
) was identified. The SmEIL1 protein was found to be localized in the nuclei, and confirmed by the transient transformation observed in tobacco leaves. The overexpression of
was able to inhibit the tanshinones accumulation to a large degree, as well as down-regulate tanshinones biosynthetic genes including
and
. These are well recognized participants in the tanshinones biosynthesis pathway. Further investigation on the
was observed to inhibit the transcription of the
gene by the Dual-Luciferase (Dual-LUC) and yeast one-hybrid (Y1H) assays. The data in this work will be of value regarding the involvement of
in regulating the biosynthesis of tanshinones and lay the foundation for the metabolic engineering of bioactive ingredients in
.
Samara oil (Elaeagnus mollis Diels kernel oil) exhibits diverse healthy functions; however, the effect of extraction on its quality is still unclear. The present study was undertaken to evaluate the ...effect of extraction methods (solvent extraction: ethyl acetate, acetone, n-hexane, and petroleum ether; mechanical extraction: hot-pressing and cold-pressing) on the color, acid value, peroxide value, fatty acid composition, bioactive compounds, antioxidant activities, and oxidative stability index of samara oil obtained from Elaeagnus mollis Diels kernels. The results indicated that extraction methods affected the physicochemical properties, chemical composition, and antioxidant activities of samara oil except for fatty acid composition and γ-tocopherol. The highest values of bioactive compounds including polyphenols (140.27 mg gallic acid equivalent (GAE)/kg) and carotenoids (42.95 mg/kg) were found in samara oil extracted with acetone. The values of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays, as well as oxidative stability index (OSI), were the highest in this oil. Correlation analysis results demonstrated that DPPH, ABTS, and OSI of samara oil were positively correlated with polyphenols and carotenoids. After evaluation, acetone could be used to extract samara oil. The study provides new information on the samara oil process.
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